Design optimization and characterization of a dual-reporter Nipah virus pseudovirus.
4 affiliations
- 1College of Animal Science, Fujian Agriculture and Forestry University, Fujian 350002, China.
- 2Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai 200241, China.
- 3School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China.
- 4Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, Jiangsu, China.
Abstract
Nipah virus (NiV), a highly pathogenic agent causing severe neurologic and respiratory disease, requires Biosafety Level 4 (BSL-4) containment. While existing pseudovirus models (e.g., based on human immunodeficiency virus (HIV) or vesicular stomatitis virus (VSV) backbones) offer alternatives, their utility is often limited by reliance on a single reporter gene. Here, we developed and optimized a robust NiV pseudovirus system utilizing a VSV-ΔG backbone co-expressing dual reporters, Green Fluorescent Protein (GFP) and NanoLuciferase (NLuc). Preliminary characterization showed that VSV-ΔG-GFP/NLuc exhibited structural integrity, achieved infection rates up to 70% in susceptible cells, and demonstrated relative stability within five freeze-thaw cycles. The study on the correlation between the expression of ephrin-B2 (EFNB2) in various cell lines and the infection efficiency of pseudovirus indicates the effectiveness of the pseudovirus. Furthermore, the platform reliably detected dose-dependent neutralization by established monoclonal antibodies targeting the fusion protein and glycoprotein of NiV, yielding trends comparable to authentic virus neutralization. Collectively, these findings establish this dual-reporter NiV pseudovirus as a safe, scalable, and high-throughput tool for antiviral screening and mechanistic studies under BSL-2 conditions. The synergistic combination of GFP (enabling visual quantification and sorting) and NLuc (providing ultra-sensitive, broad dynamic range quantification) enhances both versatility and data reliability. This study not only provides a critical new tool for NiV research but also serves as a valuable reference for developing pseudovirus platforms targeting other enveloped viruses.
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BibTeX
@article{tian2026design,
title = {{Design optimization and characterization of a dual-reporter Nipah virus pseudovirus.}},
author = {Yufei Tian and Ruixin Kuang and Ruijuan Xu and Yang Qu and Ying Liao and Cuiping Song and Lei Tan and Chan Ding and Xusheng Qiu and Yingjie Sun},
journal = {Biophysics reports},
year = {2026},
month = oct,
doi = {10.52601/bpr.2025.250063},
url = {https://europepmc.org/article/MED/42997077},
note = {CC BY},
}
RIS
TY - JOUR
TI - Design optimization and characterization of a dual-reporter Nipah virus pseudovirus.
AU - Yufei Tian
AU - Ruixin Kuang
AU - Ruijuan Xu
AU - Yang Qu
AU - Ying Liao
AU - Cuiping Song
AU - Lei Tan
AU - Chan Ding
AU - Xusheng Qiu
AU - Yingjie Sun
PY - 2026
DA - 2026/10/31
JO - Biophysics reports
DO - 10.52601/bpr.2025.250063
UR - https://europepmc.org/article/MED/42997077
AB - Nipah virus (NiV), a highly pathogenic agent causing severe neurologic and respiratory disease, requires Biosafety Level 4 (BSL-4) containment. While existing pseudovirus models (e.g., based on human immunodeficiency virus (HIV) or vesicular stomatitis virus (VSV) backbones) offer alternatives, their utility is often limited by reliance on a single reporter gene. Here, we developed and optimized a robust NiV pseudovirus system utilizing a VSV-ΔG backbone co-expressing dual reporters, Green Fluorescent Protein (GFP) and NanoLuciferase (NLuc). Preliminary characterization showed that VSV-ΔG-GFP/NLuc exhibited structural integrity, achieved infection rates up to 70% in susceptible cells, and demonstrated relative stability within five freeze-thaw cycles. The study on the correlation between the expression of ephrin-B2 (EFNB2) in various cell lines and the infection efficiency of pseudovirus indicates the effectiveness of the pseudovirus. Furthermore, the platform reliably detected dose-dependent neutralization by established monoclonal antibodies targeting the fusion protein and glycoprotein of NiV, yielding trends comparable to authentic virus neutralization. Collectively, these findings establish this dual-reporter NiV pseudovirus as a safe, scalable, and high-throughput tool for antiviral screening and mechanistic studies under BSL-2 conditions. The synergistic combination of GFP (enabling visual quantification and sorting) and NLuc (providing ultra-sensitive, broad dynamic range quantification) enhances both versatility and data reliability. This study not only provides a critical new tool for NiV research but also serves as a valuable reference for developing pseudovirus platforms targeting other enveloped viruses.
SN - 2364-3439
SN - 2364-3420
ER -